Immobilization of alkaline phosphatase using chitosan nanoparticles for enhancing its stability

Immobilization of alkaline phosphatase using chitosan nanoparticles for enhancing its stability Enzyme immobilization is an operative method for improving enzyme properties. There are various supports to enzyme immobilization. In the present study chitosan nanoparticles have been applied to alkaline phosphatase (ALP) immobilization due to their special properties. Firstly, the ionic gelation method was used to synthesize chitosan nanoparticles, which were characterized using XRD and Zetasizer Nano. Then, chitosan nanoparticles were activated with 1-ethyl-3-(3-dimethylaminopropyl carbodiimide (EDC) and ALP was successfully immobilized on them. The influence of reaction time, pH, and concentration of enzyme and EDC on efficiency of immobilization has been investigated. The studied stability against organic solvents and high temperature indicated that the stability of immobilized enzyme was increased during immobilization. The surface topography of free nanoparticles and nanoparticles with immobilized enzyme was studied by AFM. The calculation of binding efficiency showed that the immobilized ALP has conserved 71% of its native activity. The improvement of enzyme stability by immobilization indicated that chitosan nanoparticles were suitable support for immobilization. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Applied Chemistry Springer Journals

Immobilization of alkaline phosphatase using chitosan nanoparticles for enhancing its stability

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Publisher
Pleiades Publishing
Copyright
Copyright © 2014 by Pleiades Publishing, Ltd.
Subject
Chemistry; Chemistry/Food Science, general; Industrial Chemistry/Chemical Engineering
ISSN
1070-4272
eISSN
1608-3296
D.O.I.
10.1134/S107042721411024X
Publisher site
See Article on Publisher Site

Abstract

Enzyme immobilization is an operative method for improving enzyme properties. There are various supports to enzyme immobilization. In the present study chitosan nanoparticles have been applied to alkaline phosphatase (ALP) immobilization due to their special properties. Firstly, the ionic gelation method was used to synthesize chitosan nanoparticles, which were characterized using XRD and Zetasizer Nano. Then, chitosan nanoparticles were activated with 1-ethyl-3-(3-dimethylaminopropyl carbodiimide (EDC) and ALP was successfully immobilized on them. The influence of reaction time, pH, and concentration of enzyme and EDC on efficiency of immobilization has been investigated. The studied stability against organic solvents and high temperature indicated that the stability of immobilized enzyme was increased during immobilization. The surface topography of free nanoparticles and nanoparticles with immobilized enzyme was studied by AFM. The calculation of binding efficiency showed that the immobilized ALP has conserved 71% of its native activity. The improvement of enzyme stability by immobilization indicated that chitosan nanoparticles were suitable support for immobilization.

Journal

Russian Journal of Applied ChemistrySpringer Journals

Published: Mar 3, 2015

References

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